Peptide Analogs for Treating Diseases and Disorders

ABSTRACT

Provided herein are methods for the treatment of type I diabetes, Type II diabetes, metabolic syndrome, or obesity, or of appetite suppression, or for mitigating insulin resistance, or for reducing an undesirably high fasting serum glucose level, or for reducing an undesirably high peak serum glucose level, or for reducing an undesirably high peak serum insulin level, or for reducing an undesirably large response to a glucose tolerance test in synergistic combination with metformin. A peptide selected from sequences SEQ ID NO: 12, SEQ ID NO: 15 and SEQ ID NO: 17 are administered.

Cross-Reference to Related Applications

This application is a continuation under 35 U.S.C. § 120 of pendingapplication U.S. Ser. No. 15/283,338, filed Oct. 1, 2016, which is acontinuation under 35 U.S.C. § 120 of application U.S. Ser. No.14/634,188, filed Feb. 27, 2015, now U.S. Pat. No. 9,533,022, which is acontinuation-in-part under 35 U.S.C. § 120 of non-provisionalapplication U.S. Ser. No. 13/667,578, filed Nov. 2, 2012, now U.S. Pat.No. 9,006,172, which claims benefit of priority under 35 U.S.C. § 119(e)to provisional application U.S. Ser. No. 61/578,620, filed Dec. 21,2011, now abandoned, and to provisional application U.S. Ser. No.61/554,771, filed Nov. 2, 2011, the entirety of all of which are herebyincorporated herein by reference.

BACKGROUND OF THE INVENTION Field of the Invention

The embodiments disclosed herein relate to mimetics of calcitonin, andmore particularly to their use in the treatment of various diseases anddisorders, including, but not limited to diabetes (Type I and Type II),excess bodyweight, excessive food consumption and metabolic syndrome,the regulation of blood glucose levels, the regulation of response toglucose tolerance tests, the regulation of food intake, the treatment ofosteoporosis and the treatment of osteoarthritis.

Description of the Related Art

Worldwide, there are about 250 million diabetics and the number isprojected to double in the next two decades. Over 90% of this populationsuffers from type 2 diabetes mellitus (T2DM). It is estimated that only50-60% of persons affected with T2DM or in stages preceding overt T2DMare currently diagnosed. T2DM is a heterogeneous disease characterizedby abnormalities in carbohydrate and fat metabolism. The causes of T2DMare multi-factorial and include both genetic and environmental elementsthat affect β-cell function and insulin sensitivity in tissues such asmuscle, liver, pancreas and adipose tissue. As a consequence impairedinsulin secretion is observed and paralleled by a progressive decline inβ-cell function and chronic insulin resistance. The inability of theendocrine pancreas to compensate for peripheral insulin resistance leadsto hyperglycaemia and onset of clinical diabetes. Tissue resistance toinsulin-mediated glucose uptake is now recognized as a majorpathophysiologic determinant of T2DM. Type I diabetes is characterisedby a loss of the ability to produce insulin in response to food intakeand hence an inability to regulate blood glucose to a normalphysiological level.

A successful criterion for an optimal T2DM intervention is the loweringof blood glucose levels, which can be both chronic lowering of bloodglucose levels and increased ability to tolerate high glucose levelsafter food intake, described by lower peak glucose levels and fasterclearance. Both of these situations exert less strain on β-cell insulinoutput and function.

The physical structure of bone may be compromised by a variety offactors, including disease and injury. One of the most common bonediseases is osteoporosis, which is characterized by low bone mass andstructural deterioration of bone tissue, leading to bone fragility andan increased susceptibility to fractures, particularly of the hip, spineand wrist. Osteoporosis develops when there is an imbalance such thatthe rate of bone resorption exceeds the rate of bone formation.Administering an effective amount of an anti-resorptive agent, such ascalcitonin, has shown to prevent resorption of bone.

Inflammatory or degenerative diseases, including diseases of the joints,e.g. osteoarthritis (OA), rheumatoid arthritis (RA) or juvenilerheumatoid arthritis (JRA), and including inflammation that results fromautoimmune response, e.g. lupus, ankylosing spondylitis (AS) or multiplesclerosis (MS), can lead to substantial loss of mobility due to pain andjoint destruction. Cartilage that covers and cushions bone within jointsmay become degraded over time thus undesirably permitting direct contactof two bones that can limit motion of one bone relative to the otherand/or cause damage to one by the other during motion of the joint.Subchondral bone just beneath the cartilage may also degrade.Administering an effective amount of an anti-resorptive agent, such ascalcitonin, may prevent resorption of bone.

SUMMARY OF THE INVENTION

Calcitonin mimetics are disclosed herein.

According to aspects illustrated herein, there is disclosed a peptidehaving a sequence selected from SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO:13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 17 and SEQ ID NO: 18.

According to aspects illustrated herein, there is disclosed a methodthat includes administering to a patient an effective amount of apeptide selected from the group consisting of: SEQ ID NO: 11, SEQ ID NO:12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 17 toaffect a weight reduction in the patient.

According to aspects illustrated herein, there is disclosed a methodthat includes administering to a patient an effective amount of apeptide selected from the group consisting of: SEQ ID NO: 11, SEQ ID NO:12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, and SEQ ID NO: 17 toaffect postprandial glycemic control in the patient.

According to aspects illustrated herein, there is disclosed a methodthat includes administering to a patient an effective amount of apeptide selected from the group consisting of: SEQ ID NO: 11, SEQ ID NO:12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, and SEQ

ID NO: 17 to affect an improvement in glycemic control in the patient.

According to aspects illustrated herein, there is disclosed a methodthat includes administering to a patient an effective amount of apeptide of SEQ ID NO: 18 having the sequenceC_(m)SNLSTCVLGKLSQELHKLQTYPRTDVGANXaaXaa_(a) so as to reduce at leastone of bone resorption and cartilage degradation in the patient.

According to aspects illustrated herein, there is disclosed a method forthe treatment of type I diabetes, Type II diabetes, metabolic syndrome,or obesity, or of appetite suppression, or for mitigating insulinresistance, or for reducing an undesirably high fasting serum glucoselevel, or for reducing an undesirably high peak serum glucose level, orfor reducing an undesirably high peak serum insulin level, or forreducing an undesirably large response to a glucose tolerance testcomprising administering to a patient as a combination therapy aneffective amount metformin and of a peptide of SEQ ID NOS: 11-24. It hasbeen found that the subject peptides show a synergistic activity whenadministered in combination with metformin.

BRIEF DESCRIPTION OF THE DRAWINGS

The presently disclosed embodiments will be further explained withreference to the attached drawings, which illustrate the principlesthereof.

FIG. 1A, FIG. 1B, FIG. 1C, and FIG. 1D show the effect of chronic oralsalmon calcitonin (“sCT”) versus oral UGP 302 administration food intake(FIG. 1A), on body weight (FIG. 1B), cumulative food intake (FIG. 1C),and change in body weight (FIG. 1D) in DIO rats as measured in Example1;

FIG. 2A and FIG. 2B show the effect of oral sCT versus oral UGP 302 onglucose tolerance during OGTT in DIO rats as measured in Example 1.

FIG. 3 shows the effect of oral sCT versus oral UGP 302 on fastingglycemia in DIO rats as measured in Example 1.

FIG. 4A and FIG. 4B show the effect of oral sCT versus oral UGP 302 onbody weight (FIG. 4A) and food intake (FIG. 4B) in DIO rats observed inExample 2 at a first dosage.

FIG. 5A and FIG. 5B show the effect of oral sCT versus oral UGP 302 onbody weight (FIG. 5A) and food intake (FIG. 5B) in DIO rats observed inExample 2 at a second dosage.

FIG. 6A and FIG. 6B show the effect of oral sCT versus oral UGP 302 onbody weight (FIG. 6A) and food intake (FIG. 6B) in DIO rats observed inExample 2 at a third dosage.

FIG. 7A and FIG. 7B show the effect of oral sCT versus oral UGP 302 at afirst dosage on glucose tolerance during OGTT in DIO rats as measured inExample 2.

FIG. 8A and FIG. 8B show the effect of oral sCT versus oral UGP 302 at asecond dosage on glucose tolerance during OGTT in DIO rats as measuredin Example 2.

FIG. 9A, FIG. 9B, FIG. 9C, FIG. 9D, FIG. 9E, and FIG. 9F show the effectof oral sCT versus three oral UGPs on body weight (FIGS. 9A, 9C, 9E) andfood intake (FIGS. 9B, 9C) in DIO rats as measured in Example 3.

FIG. 10A, FIG. 10B, FIG. 10C, FIG. 10D, FIG. 10E, and FIG. 10F show theeffect of oral sCT versus three oral UGPs on glucose levels in a glucosetolerance test in DIO rats as measured in Example 3.

FIG. 11 shows binding results for six UGP compounds to T47D cellcalcitonin receptors as measured in Example 4.

FIG. 12A and FIG. 12B show food consumption (FIG. 12A) and weight changemeasurements (FIG. 12B) for UGP 282 as measured in Example 5.

FIG. 13A and FIG. 13B show food consumption (FIG. 13A) and weight changemeasurements (FIG. 13B) for UGP 283 as measured in Example 5.

FIG. 14A and FIG. 14B show food consumption (FIG. 14A) and weight changemeasurements (FIG. 14B) for UGP 284 as measured in Example 5.

FIG. 15A and FIG. 15B show food consumption (FIG. 15A) and weight changemeasurements (FIG. 15B) for UGP 298 as measured in Example 5.

FIG. 16A and FIG. 16B show food consumption (FIG. 16A) and weight changemeasurements (FIG. 16B) for UGP 302 as measured in Example 5.

FIG. 17A and FIG. 17B show food consumption (FIG. 17A) and weight changemeasurements (FIG. 17B) for UGP 303 as measured in Example 5;

FIG. 18 shows the reduction of bone resorption produced by treatmentwith UGP302 in rats.

FIG. 19 shows cartilage resorption produced by treatment with UGP302 inrats.

FIG. 20 shows results obtained in Example 7 showing fasting bloodglucose (FBG) of tested rats.

FIG. 21 shows results obtained in Example 7 showing non-fasted bloodglucose (PPG) of subject rats.

FIG. 22 shows results obtained in Example 7 showing HbA1c levels ofsubject rats.

FIG. 23A and FIG. 23B show results obtained in Example 7 showing theresults of oral glucose tolerance tests (OGTT) as plots of blood glucoseagainst time (FIG. 23A) and as total Area Under Curve tAUC measurements(FIG. 23B).

FIG. 24A and FIG. 24B show results obtained in Example 7 showing theresults of insulin tolerance tests (IPITT) as plots of blood glucoseagainst time (FIG. 24A) and as total Area Under Curve tAUC measurements(FIG. 24B).

While the above-identified drawings set forth presently disclosedembodiments, other embodiments are also contemplated, as noted in thediscussion. This disclosure presents illustrative embodiments by way ofrepresentation and not limitation. Numerous other modifications andembodiments can be devised by those skilled in the art which fall withinthe scope and spirit of the principles of the presently disclosedembodiments.

DETAILED DESCRIPTION OF THE INVENTION

Calcitonins are highly conserved over a wide range of species.Full-length native calcitonin is 32 amino acids in length. The sequencesof examples of calcitonins are set out below:

SEQ ID NO: 1 Salmon CSNLSTCVLGKLSQELHKLQTYPRTNTGSGTP  SEQ ID NO: 2Mouse CGNLSTCMLGTYTQDLNKFHTFPQTSIGVEAP  SEQ ID NO: 3Chicken CASLSTCVLGKLSQELHKLQTYPRTDVGAGTP SEQ ID NO: 4Eel CSNLSTCVLGKLSQELHKLQTYPRTDVGAGTP SEQ ID NO: 5Rat CGNLSTCMLGTYTQDLNKFHTFPQTSIGVGAP SEQ ID NO: 6Horse CSNLSTCVLGTYTQDLNKFHTFPQTAIGVGAP SEQ ID NO: 7Canine-1 CSNLSTCVLGTYSKDLNNFHTFSGIGFGAETP SEQ ID NO: 8Canine-2 CSNLSTCVLGTYTQDLNKFHTFPQTAIGVGAP SEQ ID NO: 9Porcine CSNLSTCVLSAYWRNLNNFHRFSGMGFGPETP SEQ ID NO: 10Human CGNLSTCMLGTYTQDFNKFHTFPQTAIGVGAP

Embodiments of the present disclosure relate to calcitonin mimetics. Theamino acid sequence of the calcitonin mimetics of the present disclosureare found in Table 1A below.

TABLE 1A  Calcitonin SEQ Mimetic ID (CM) Amino Acid Sequence NO: UGP281 AcCSNLSTCVLGKLSQELHKLQTYPRTDVGA 11 NTY-NH₂ UGP283 AcCSNLSTCVLGRLSQELHRLQTFPRTDVGA 12 NTAcY UGP284 PrCSNLSTCVLGKLSQELHKLQTYPRTNTGS 13 GTP-NH₂ UGP298 SuccCSNLSTCVLGKLSQELHKLQTYPRTNT 14 GSGTP-NH₂ UGP302 AcCSNLSTCVLGKLSQELHKLQTYPRTDVGA 15 NAP-NH₂ UGP303 KCSNLSTCVLGKLSQELHKLQTYPRTDVGAN 16 TY-NH₂ UGP306 SuccCSNLSTCVLGKLSQELHKLQTYPRTDV 17 GANAY-NH₂ UGP1000C_(m)SNLSTCVLGKLSQELHKLQTYPRTDVGAN 18 XaaXaa_(a)

In some embodiments, the cysteine at position 1 of the calcitoninmimetics discussed supra is modified (“C_(m)”) to reduce the positivecharge of the first amino acid. For example, an acetyl group (SEQ IDNOs: 11, 12 and 15), propionyl group (SEQ ID NO: 13), or succinyl group(SEQ ID NOs: 14 and 17) may be substituted on cysteine-1. In someembodiments, the amino acid at the last position (“Xaa_(a)”) (position32 in SEQ ID Nos: 11, 13-15 and 17-18 or position 33 in SEQ ID NO: 16)may include an amidated group “NH₂”. Alternative ways of reducingpositive charge include, but are not limited to, polyethyleneglycol-based PEGylation, or the addition of another amino acid such asglutamic acid or aspartic acid at the N-terminus. Alternatively, otheramino acids may be added to the N-terminus of peptides discussed supraincluding, but not limited to, lysine, glycine, formylglycine, leucine,alanine, acetyl alanine, and dialanyl. An example of an amino acid addedto the N-terminus of peptides includes SEQ ID NO:16, where a lysine hasbeen added.

“Xaa” in SEQ ID NO: 18 in Table 1 can be any naturally occurring aminoacid. In an embodiment Xaa at position 31 is selected from one ofthreonine or alanine. In an embodiment Xaa at position 32 is selectedfrom one of tyrosine or proline. Thus, SEQ ID NOs: 11, 15, 16 and 17,are encompassed by SEQ ID NO: 18.

Other amino acid sequences of the calcitonin mimetics of the presentdisclosure are found in Table 1B below. In these sequences the Rsubstituent may be an acylation moiety or may be absent.

TABLE 1B  SEQ ID Amino Acid Sequence NO:R-CSNLSTCVLGKLSQELHKLQTYPRTDVGANAP-NH₂ 19R-CSNLSTCVLGKLSQELHKLQTYPRTNTGSGTP-NH₂ 20R-CSNLSTCVLGKLSQELHKLQTYPRTDVGANTY-NH₂ 21R-CSNLSTCVLGRLSQELHRLQTFPRTDVGANTAcY 22R-KCSNLSTCVLGKLSQELHKLQTYPRTDVGANTY-NH₂ 23R-CSNLSTCVLGKLSQELHKLQTYPRTDVGANAY-NH₂ 24

As those of skill in the art will appreciate, peptides having aplurality of cysteine residues frequently form a disulfide bridgebetween two such cysteine residues. All such peptides set forth hereinare defined as optionally including one or more such disulfide bridges.While calcitonin mimetics of the present disclosure may exist in freeacid form, it is preferred that the C-terminal amino acid be amidated.Applicants expect that such amidation may contribute to theeffectiveness and/or bioavailability of the peptide. A preferredtechnique for manufacturing amidated versions of the calcitonin mimeticsof the present disclosure is to react precursors (having glycine inplace of the C-terminal amino group of the desired amidated product) inthe presence of peptidylglycine alpha-amidating monooxygenase inaccordance with known techniques wherein the precursors are converted toamidated products in reactions described, for example, in U.S. Pat. No.4,708,934 and European Patent Publication Nos. 0 308 067 and 0 382 403.Recombinant production is preferred for both the precursor and theenzyme that catalyzes the conversion of the precursor to salmoncalcitonin. Such recombinant production is discussed in Biotechnology,Vol. 11 (1993) pp. 64-70, which further describes a conversion of aprecursor to an amidated product. The recombinant product reported thereis identical to natural salmon calcitonin, and to salmon calcitoninproduced using solution and solid phase chemical peptide synthesis.Production of amidated products may also be accomplished using theprocess and amidating enzyme set forth by Consalvo, et al. in U.S. Pat.No. 7,445,911; Miller et al., U.S. Patent Publication No. 2006/0292672;Ray et al. 2002, Protein Expression and Purification, 26:249-259; andMehta, 2004, Biopharm. International, July, pp. 44-46.

The production of the preferred amidated peptides may proceed, forexample, by producing glycine-extended precursor in E. coli as a solublefusion protein with glutathione-S-transferase, or by direct expressionof the precursor in accordance with the technique described in U.S. Pat.No. 6,103,495. Such a glycine extended precursor has a molecularstructure that is identical to the desired amidated product except atthe C-terminus (where the product terminates —X—NH₂, while the precursorterminates —X-gly, X being the C-terminal amino acid residue of theproduct). An alpha-amidating enzyme described in the publications abovecatalyzes conversion of precursors to product. That enzyme is preferablyrecombinantly produced, for example, in Chinese Hamster Ovary (CHO)cells), as described in the Biotechnology and Biopharm. articles citedabove.

Free acid forms of peptide active agents of the present disclosure maybe produced in like manner, except without including a C-terminalglycine on the “precursor”, which precursor is instead the final peptideproduct and does not require the amidation step.

Except where otherwise stated, the preferred dosage of the calcitoninmimetics of the present disclosure is identical for both therapeutic andprophylactic purposes. Desired dosages are discussed in more detail,infra, and differ depending on mode of administration.

Except where otherwise noted or where apparent from context, dosagesherein refer to weight of active compounds unaffected by pharmaceuticalexcipients, diluents, carriers or other ingredients, although suchadditional ingredients are desirably included, as shown in the examplesherein. Any dosage form (capsule, tablet, injection or the like)commonly used in the pharmaceutical industry for delivery of peptideactive agents is appropriate for use herein, and the terms “excipient”,“diluent”, or “carrier” includes such non-active ingredients as aretypically included, together with active ingredients in such dosage formin the industry. A preferred oral dosage form is discussed in moredetail, infra, but is not to be considered the exclusive mode ofadministering the active agents of the present disclosure.

The calcitonin mimetics of the present disclosure can be administered toa patient to treat a number of diseases or disorders. As used herein,the term “patient” means any organism belonging to the kingdom Animalia.In an embodiment, the term “patient” refers to vertebrates, morepreferably, mammals including humans.

Accordingly, the present disclosure provides a method of treatment oftype I diabetes, Type II diabetes or metabolic syndrome, obesity, or ofappetite suppression, or for mitigating insulin resistance, or forreducing an undesirably high fasting serum glucose level, or forreducing an undesirably high peak serum glucose level, or for reducingan undesirably high peak serum insulin level, or for reducing anundesirably large response to a glucose tolerance test, or for treatingosteoporosis, or for treating osteoarthritis.

As used herein, the term “glycemic control” refers to the typical levelsof blood sugar (glucose)in a person with diabetes mellitus. Thepercentage of hemoglobin which is glycosolated (measured as hemoglobinA1c) is used as a proxy measure of long-term glycemic control.

As used herein, the term “improved glycemic control” refers to theability of a calcitonin mimetic of the present disclosure to reduce thepercentage of hemoglobin which is glycosolated.

There are a number of art-recognized measures of normal range for bodyweight in view of a number of factors such as gender, age and height. Apatient in need of treatment or prevention regimens set forth hereininclude patients whose body weight exceeds recognized norms or who, dueto heredity, environmental factors or other recognized risk factor, areat higher risk than the general population of becoming overweight orobese. In accordance with the present disclosure, it is contemplatedthat the calcitonin mimetics may be used to treat diabetes where weightcontrol is an aspect of the treatment.

In an embodiment, the method includes enteral administration to apatient in need thereof for treatment of a said condition of apharmaceutically effective amount of any one of the peptides describedherein.

In an embodiment, the method includes parenteral administration to apatient in need thereof for treatment of a said condition of apharmaceutically effective amount of any one of the peptides describedherein. For parenteral administration (including intraperitoneal,subcutaneous, intravenous, intradermal or intramuscular injection),solutions of a peptide of the present disclosure in either sesame orpeanut oil or in aqueous propylene glycol may be employed, for example.The aqueous solutions should be suitably buffered (preferably pH greaterthan 8) if necessary and the liquid diluent first rendered isotonic.These aqueous solutions are suitable for intravenous injection purposes.The oily solutions are suitable for intraarticular, intramuscular andsubcutaneous injection purposes. The preparation of all these solutionsunder sterile conditions is readily accomplished by standardpharmaceutical techniques well known to those skilled in the art. Forparenteral application, examples of suitable preparations includesolutions, preferably oily or aqueous solutions as well as suspensions,emulsions, or implants, including suppositories. Peptides may beformulated in sterile form in multiple or single dose formats such asbeing dispersed in a fluid carrier such as sterile physiological salineor 5% saline dextrose solutions commonly used with injectables.

Said method may include a preliminary step of determining whether thepatient suffers from a said condition, and/or a subsequent step ofdetermining to what extent said treatment is effective in mitigating thecondition in said patient, e.g. in each case, carrying out an oralglucose tolerance test or a resting blood sugar level.

For improved control over the weight of the patient, to produce a lossof weight or an avoidance of weight gain, the active compound ispreferably administered at least twice per day, e.g. from 2-4 times perday. Formulations of the active compound may contain a unit dosageappropriate for such an administration schedule. The active compoundsmay be administered with a view to controlling the weight of a patientundergoing treatment for diabetes or metabolic syndrome.

Oral enteral formulations are for ingestion by swallowing for subsequentrelease in the intestine below the stomach, and hence delivery via theportal vein to the liver, as opposed to formulations to be held in themouth to allow transfer to the bloodstream via the sublingual or buccalroutes.

Suitable dosage forms for use in the present disclosure include tablets,mini-tablets, capsules, granules, pellets, powders, effervescent solidsand chewable solid formulations. Such formulations may include gelatinwhich is preferably hydrolysed gelatin or low molecular weight gelatin.Such formulations may be obtainable by freeze drying a homogeneousaqueous solution comprising calcitonin or a fragment or conjugatethereof and hydrolysed gelatin or low molecular weight gelatin andfurther processing the resulting solid material into said oralpharmaceutical formulation, and wherein the gelatin may have a meanmolecular weight from 1000 to 15000 Daltons. Such formulations mayinclude a protective carrier compound such as 5-CNAC or others asdisclosed herein.

Whilst oral formulations such as tablets and capsules are preferred,compositions for use in the present disclosure may take the form ofsyrups, elixirs or the like and suppositories or the like. Oral deliveryis generally the delivery route of choice since it is convenient,relatively easy and generally painless, resulting in greater patientcompliance relative to other modes of delivery. However, biological,chemical and physical barriers such as varying pH in thegastrointestinal tract, powerful digestive enzymes, and active agentimpermeable gastrointestinal membranes, makes oral delivery ofcalcitonin like peptides to mammals problematic, e.g. the oral deliveryof calcitonins, which are long-chain polypeptide hormones secreted bythe parafollicular cells of the thyroid gland in mammals and by theultimobranchial gland of birds and fish, originally proved difficultdue, at least in part, to the insufficient stability of calcitonin inthe gastrointestinal tract as well as the inability of calcitonin to bereadily transported through the intestinal walls into the blood stream.

Suitable oral formulations are however described below.

Treatment of Patients

In an embodiment, a calcitonin mimetic of the present disclosure isadministered at adequate dosage to maintain serum levels of the mimeticin patients between 5 and 500 picograms per milliliter, preferablybetween 10 and 250 picograms per milliliter. The serum levels may bemeasured by radioimmunoassay techniques known in the art. The attendingphysician may monitor patient response, and may then alter the dosagesomewhat to account for individual patient metabolism and response. Nearsimultaneous release is best achieved by administering all components ofthe present disclosure as a single pill or capsule. However, thedisclosure also includes, for example, dividing the required amount ofthe calcitonin mimetic among two or more tablets or capsules which maybe administered together such that they together provide the necessaryamount of all ingredients. “Pharmaceutical composition,” as used hereinincludes but is not limited to a complete dosage appropriate to aparticular administration to a patient regardless of whether one or moretablets or capsules (or other dosage forms) are recommended at a givenadministration.

A calcitonin mimetic of the present disclosure may be formulated fororal administration using the methods employed in the Unigene Enteripep®products. These may include the methods as described in U.S. Pat. No.5,912,014, U.S. Pat. No. 6,086,918, U.S. Pat. No. 6,673,574, U.S. Pat.No. 7,316,819, U.S. Pat. No. 8,093,207, and US Publication No.2009/0317462. In particular, it may include the use of conjugation ofthe compound to a membrane translocator such as the protein transductiondomain of the HIV TAT protein, co-formulation with one or more proteaseinhibitors, and/or a pH lowering agent which may be coated and/or anacid resistant protective vehicle and/or an absorption enhancer whichmay be a surfactant.

In an embodiment, a calcitonin mimetic of the present disclosure ispreferably formulated for oral delivery in a manner known in U.S. PatentPublication No. 2009/0317462. One preferred oral dosage form inaccordance with the present disclosure is set forth in Table 2 below:

TABLE 2 ACTIVE AGENT OR EXCIPIENT FUNCTION A Calcitonin Mimetic selectedfrom Active agent one of SEQ ID NO: 11 SEQ ID NO: 18 Coated Citric AcidParticles Protease Inhibitor Lauroylcarnitine Absorption EnhancerNonionic Polymer Subcoat Eudragit L30D-55 Enteric Coat

In an embodiment, a calcitonin mimetic of the present disclosure may beformulated for enteral, especially oral, administration by admixturewith a suitable carrier compound. Suitable carrier compounds includethose described in U.S. Pat. No. 5,773,647 and U.S. Pat. No. 5,866,536and amongst these, 5-CNAC (N-(5-chlorosalicyloyl)-8-aminocaprylic acid,commonly as its disodium salt) is particularly effective. Otherpreferred carriers or delivery agents are SNAD (sodium salt of10-(2-Hydroxybenzamido)decanoic acid) and SNAC (sodium salt ofN-(8-[2-hydroxybenzoyl]amino)caprylic acid). In an embodiment, apharmaceutical composition of the present disclosure comprises adelivery effective amount of carrier such as 5-CNAC, i.e. an amountsufficient to deliver the compound for the desired effect. Generally,the carrier such as 5-CNAC is present in an amount of 2.5% to 99.4% byweight, more preferably 25% to 50% by weight of the total composition.

In addition, WO 00/059863 discloses the disodium salts of formula I

wherein R¹, R², R³, and R⁴ are independently hydrogen, —OH, —NR⁶R⁷,halogen, C₁-C₄ alkyl, or C₁-C₄ alkoxy; R⁵ is a substituted orunsubstituted C₂-C₁₆ alkylene, substituted or unsubstituted C₂-C₁₆alkenylene, substituted or unsubstituted C₁-C₁₂ alkyl(arylene), orsubstituted or unsubstituted aryl(C₁-C₁₂ alkylene); and R⁶ and R⁷ areindependently hydrogen, oxygen, or C₁-C₄ alkyl; and hydrates andsolvates thereof as particularly efficacious for the oral delivery ofactive agents, such as calcitonins, e.g. salmon calcitonin, and thesemay be used in the present disclosure.

Preferred enteric formulations using optionally micronised 5-CNAC may begenerally as described in WO2005/014031.

The compound may be formulated for oral administration using the methodsemployed in the Capsitonin product of Bone Medical Limited. These mayinclude the methods incorporated in Axcess formulations. Moreparticularly, the active ingredient may be encapsulated in an entericcapsule capable of withstanding transit through the stomach. This maycontain the active compound together with a hydrophilic aromatic alcoholabsorption enhancer, for instance as described in WO02/028436. In aknown manner the enteric coating may become permeable in a pH sensitivemanner, e.g. at a pH of from 3 to 7. WO2004/091584 also describessuitable formulation methods using aromatic alcohol absorptionenhancers.

The compound may be formulated using the methods seen in the Oramedproducts, which may include formulation with omega-3 fatty acid as seenin WO2007/029238 or as described in U.S. Pat. No. 5,102,666.

Generally, the pharmaceutically acceptable salts (especially mono or disodium salts), solvates (e.g. alcohol solvates) and hydrates of thesecarriers or delivery agents may be used.

Oral administration of the pharmaceutical compositions according to thedisclosure can be accomplished regularly, e.g. once or more on a dailyor weekly basis; intermittently, e.g.

irregularly during a day or week; or cyclically, e.g. regularly for aperiod of days or weeks followed by a period without administration. Thedosage form of the pharmaceutical compositions of the presentlydisclosed embodiments can be any known form, e.g. liquid or solid dosageforms. The liquid dosage forms include solution emulsions, suspensions,syrups and elixirs. In addition to the active compound and carrier suchas 5-CNAC, the liquid formulations may also include inert excipientscommonly used in the art such as, solubilizing agents e.g. ethanol; oilssuch as cottonseed, castor and sesame oils; wetting agents; emulsifyingagents; suspending agents; sweeteners; flavourings; and solvents such aswater. The solid dosage forms include capsules, soft-gel capsules,tablets, caplets, powders, granules or other solid oral dosage forms,all of which can be prepared by methods well known in the art. Thepharmaceutical compositions may additionally comprise additives inamounts customarily employed including, but not limited to, a pHadjuster, a preservative, a flavorant, a taste-masking agent, afragrance, a humectant, a tonicifier, a colorant, a surfactant, aplasticizer, a lubricant such as magnesium stearate, a flow aid, acompression aid, a solubilizer, an excipient, a diluent such asmicrocrystalline cellulose, e.g. Avicel PH 102 supplied by FMCcorporation, or any combination thereof. Other additives may includephosphate buffer salts, citric acid, glycols, and other dispersingagents. The composition may also include one or more enzyme inhibitors,such as actinonin or epiactinonin and derivatives thereof; aprotinin,Trasylol and Bowman-Birk inhibitor.

Further, a transport inhibitor, i.e. a [rho]-glycoprotein such asKetoprofin, may be present in the compositions of the presentdisclosure. The solid pharmaceutical compositions of the instantdisclosure can be prepared by conventional methods e.g. by blending amixture of the active compound, the carrier such as 5-CNAC, and anyother ingredients, kneading, and filling into capsules or, instead offilling into capsules, molding followed by further tableting orcompression-molding to give tablets. In addition, a solid dispersion maybe formed by known methods followed by further processing to form atablet or capsule. Preferably, the ingredients in the pharmaceuticalcompositions of the instant disclosure are homogeneously or uniformlymixed throughout the solid dosage form.

Alternatively, the active compound may be formulated as a conjugate withsaid carrier, which may be an oligomer as described in US2003/0069170,e.g.

Such conjugates may be administered in combination with a fatty acid anda bile salt as described there.

Conujugates with polyethylene glycol (PEG) may be used, as described forinstance in Mansoor et al.

Alternatively, active compounds may be admixed withnitroso-N-acetyl-D,L-penicillamine (SNAP) and Carbopol solution or withtaurocholate and Carbapol solution to form a mucoadhesive emulsion.

The active compound may be formulated by loading into chitosannanocapsules as disclosed in Prego et al. (optionally PEG modified as inPrego Prego C, Torres D, Fernandez-Megia E, Novoa-Carballal R, Quiñoá E,Alonso M J.) or chitosan or PEG coated lipid nanoparticles as disclosedin Garcia-Fuentes et al. Chitosan nanoparticles for this purpose may beiminothiolane modified as described in Guggi et al. They may beformulated in water/oil/water emulsions as described in Dogru et al. Thebioavailability of active compounds may be increased by the use oftaurodeoxycholate or lauroyl carnitine as described in Sinko et al. orin Song et al. Generally, suitable nanoparticles as carriers arediscussed in de la Fuente et al and may be used in the presentdisclosure.

Other suitable strategies for oral formulation include the use of atransient permeability enhancer (TPE) system as described inWO2005/094785 of Chiasma Ltd. TPE makes use of an oily suspension ofsolid hydrophilic particles in a hydrophobic medium to protect the drugmolecule from inactivation by the hostile gastrointestinal (GI)environment and at the same time acts on the GI wall to inducepermeation of its cargo drug molecules.

Further included is the use of glutathione or compounds containingnumerous thiol groups as described in US2008/0200563 to inhibit theaction of efflux pumps on the mucous membrane. Practical examples ofsuch techniques are described also in Caliceti, P. Salmaso, S., Walker,G. and Bernkop-Schnürch, A. (2004) ‘Development and in vivo evaluationof an oral insulin-PEG delivery system.’ Eur. J. Pharm. Sci., 22,315-323, in Guggi, D., Krauland, A. H., and Bernkop-Schnurch, A. (2003)‘Systemic peptide delivery via the stomach: in vivo evaluation of anoral dosage form for salmon calcitonin’. J. Control. Rel. 92,125-135,and in Bernkop-Schnürch, A., Pinter, Y., Guggi, D., Kahlbacher, H.,Schöffmann, G., Schuh, M., Schmerold, I., Del Curto, M.D., D'Antonio,M., Esposito, P. and Huck, Ch. (2005) ‘The use of thiolated polymers ascarrier matrix in oral peptide delivery’—Proof of concept. J. Control.Release, 106, 26-33.

The active compound may be formulated in seamless micro-spheres asdescribed in WO2004/084870 where the active pharmaceutical ingredient issolubilised as an emulsion, microemulsion or suspension, formulated intomini-spheres; and variably coated either by conventional or novelcoating technologies. The result is an encapsulated drug in“pre-solubilised” form which when administered orally provides forpredetermined instant or sustained release of the active drug tospecific locations and at specific rates along the gastrointestinaltract. In essence, pre-solubilization of the drug enhances thepredictability of its kinetic profile while simultaneously enhancingpermeability and drug stability.

One may employ chitosan coated nanocapsules as described inUS2009/0074824. The active molecule administered with this technology isprotected inside the nanocapsules since they are stable against theaction of the gastric fluid. In addition, the mucoadhesive properties ofthe system enhances the time of adhesion to the intestine walls (it hasbeen verified that there is a delay in the gastrointestinal transit ofthese systems) facilitating a more effective absorption of the activemolecule.

Methods developed by TSRI Inc. may be used. These include HydrophilicSolubilization Technology (HST) in which gelatin, a naturally derivedcollagen extract carrying both positive and negative charges, coats theparticles of the active ingredient contained in lecithin micelles andprevents their aggregation or clumping. This results in an improvedwettability of hydrophobic drug particles through polar interactions. Inaddition, the amphiphilic lecithin reduces surface tension between thedissolution fluid and the particle surface.

The active ingredient may be formulated with cucurbiturils asexcipients.

Alternatively, one may employ the GIPET technology of MerrionPharmaceuticals to produce enteric coated tablets containing the activeingredient with an absorption enhancer which may be a medium chain fattyacid or a medium chain fatty acid derivative as described inUS2007/0238707 or a membrane translocating peptide as described in U.S.Pat. No. 7,268,214.

One may employ GIRES™ technology which consists of a controlled-releasedosage form inside an inflatable pouch, which is placed in a drugcapsule for oral administration. Upon dissolution of the capsule, agas-generating system inflates the pouch in the stomach. In clinicaltrials the pouch has been shown to be retained in the stomach for 16-24hours.

Alternatively, the active may be conjugated to a protective modifierthat allows it to withstand enzymatic degradation in the stomach andfacilitate its absorption. The active may be conjugated covalently witha monodisperse, short-chain methoxy polyethylene glycol glycolipidsderivative that is crystallized and lyophilized into the dry activepharmaceutical ingredient after purification. Such methods are describedin U.S. Pat. No. 5,438,040.

One may also employ a hepatic-directed vesicle (HDV) for activedelivery. An HDV may consist of liposomes (150 nm diameter)encapsulating the active, which also contain a hepatocyte-targetingmolecule in their lipid bilayer. The targeting molecule directs thedelivery of the encapsulated active to the liver cells and thereforerelatively minute amounts of active are required for effect. Suchtechnology is described in US2009/0087479.

The active may be incorporated into a composition containingadditionally a substantially non-aqueous hydrophilic medium comprisingan alcohol and a cosolvent, in association with a medium chain partialglyceride, optionally in admixture with a long-chain PEG species asdescribed in US2002/0115592 in relation to insulin.

Alternatively, use may be made of intestinal patches as described inShen Z, Mitragotri S, Pharm Res. 2002 Apr;19(4):391-5 ‘Intestinalpatches for oral drug delivery’.

The active may be incorporated into an erodible matrix formed from ahydrogel blended with a hydrophobic polymer as described in U.S. Pat.No. 7,189,414.

Suitable oral dosage levels for adult humans to be treated may be in therange of 0.05 to 5 mg, preferably about 0.1 to 2.5mg.

The frequency of dosage treatment of patients may be from 1 to six timesdaily, for instance from two to four times daily. Treatment willdesirably be maintained over a prolonged period of at least 6 weeks,preferably at least 6 months, preferably at least a year, and optionallyfor life.

Combination treatments for relevant conditions may be carried out usinga composition according to the present disclosure and separateadministration of one or more other therapeutics. Alternatively, thecomposition according to the present disclosure may incorporate one ormore other therapeutics for combined administration.

Combination therapies according to the present disclsoure includecombinations of an active compound as described with insulin, GLP-2,GLP-1, GIP, or amylin, or generally with other anti-diabetics. Thuscombination therapies including co-formulations may be made with insulinsensitizers including biguanides such as Metformin, Buformin andPhenformin, TZD's (PPAR) such as Balaglitazone, Pioglitazone,Rivoglitazone, Rosiglitazone and Troglitazone, dual PPAR agonists suchas Aleglitazar, Muraglitazar and Tesaglitazar, or secretagoguesincluding sulphonylureas such as Carbutamide, Chloropropamide,Gliclazide, Tolbutamide, Tolazamide, Glipizide, Glibenclamide,Glyburide, Gliquidone, Glyclopyramide and Glimepriride,Meglitinides/glinides (K+) such as Nateglinide, Repaglinide andMitiglinide, GLP-1 analogs such as Exenatide, Liraglutide andAlbiglutide, DPP-4 inhibitors such as Alogliptin, Linagliptin,Saxagliptin, Sitagliptin and Vildagliptin, insulin analogs or specialformulations such as (fast acting) Insulin lispro, Insulin aspart,Insulin glulisine, (long acting) Insulin glargine, Insulin detemir),inhalable insulin—Exubra and NPH insulin, and others includingalpha-glucosidase inhibitors such as Acarbose, Miglitol and Voglibose,amylin analogues such as Pramlintide, SGLT2 inhibitors such asDapagliflozin, Remogliflozin and Sergliflozin as well as miscellaneousones including Benfluorex and Tolrestat.

Further combinations include co-administration or co-formulation withleptins. Leptin resistance is a well-established component of type 2diabetes; however, injections of leptin have so far failed to improveupon this condition. In contrast, there is evidence supporting thatamylin, and thereby molecules with amylin-like abilities, as the salmoncalcitonin mimetics, are able to improve leptin sensitivity.Amylin/leptin combination has shown a synergistic effect on body weightand food intake, and also insulin resistance [Kusakabe T et al].Accordingly, the present disclosure provides a compound of the formulaAc-CSNLSTCVLG KLSQELHKLQ TYPRTDVGAN AP-NH₂ (SEQ ID NO: 15), which willbe referred to herein as ‘calcitonin mimetic 1’ or ‘UGP302’.

Accordingly, the present disclosure includes a pharmaceuticalformulation of such a peptide for enteral administration, e.g. fortreating type I diabetes, type II diabetes, or metabolic syndrome, orfor mitigating insulin resistance, or for reducing an undesirably highfasting serum glucose level, or for reducing an undesirably high peakserum glucose level, or for reducing an undesirably high peak seruminsulin level, or for reducing an undesirably high response to a glucosetolerance test, or for treating osteoporosis, or for treatingosteoarthritis. The formulation may comprise also a carrier serving toenable effective enteral administration of said active compound.

Preferably, said formulation is formulated for oral administration tothe digestive tract.

Preferably, said carrier comprises 5-CNAC, SNAD, or SNAC.

Additionally, the present disclosure includes said peptides as newcompounds.

The presently disclosed embodiments is described in the followingExamples, which are set forth to aid in the understanding of thedisclosure, and should not be construed to limit in any way the scope ofthe disclosure as defined in the claims which follow thereafter. Thefollowing examples are put forth so as to provide those of ordinaryskill in the art with a complete disclosure and description of how tomake and use the described embodiments, and are not intended to limitthe scope of the present disclosure nor are they intended to representthat the experiments below are all or the only experiments performed.Efforts have been made to ensure accuracy with respect to numbers used(e.g. amounts, temperature, etc.) but some experimental errors anddeviations should be accounted for. Unless indicated otherwise, partsare parts by weight, molecular weight is weight average molecularweight, temperature is in degrees Centigrade, and pressure is at or nearatmospheric.

EXAMPLE 1 Chronic Effect of Calcitonin Mimetic 1 (CM1) Compared to sCTAnimals

The study was performed in male Levin-DIO rats (diet-sensitive) andLevin-DR (diet-resistant) (TacLevin: CD (SD) DIO) (Taconic, Hudson,N.Y., U.S.A.) obtained at age 6-7 weeks.

On arrival, DIO rats were given high fat diet (60 kcal %) (#D12495,Research Diets Inc., New Brunswick, N.J., USA) and kept on the same dietfor 16 weeks prior to and during the experiment. DR rats were givenlow-fat diet and served as control group. Animals were pair-wise housedthroughout the study. Rats were handled and pre-dosed once daily withMilliQ H20 for 2-3 weeks prior to experimental start to reducestress-induced hyperglycaemia. Baseline parameters were recorded in anfasting (6 h) condition. Rats were randomized into treatment groupsbased on fasting body weight (BW) and fasting plasma glucose (FPG). Bodyweight, food and water intake were recorded once weekly during the studyperiod.

Compound

Oral sCT or calcitonin mimetic 1 solution was prepared on the day ofdosing by mixing a carrier with the given compound based on thefollowing calculations:

5-CNAC (vehicle):

Animals treated with oral 5-CNAC received a dose of 150 mg/kg dissolvedin milliQ H2O.

-   Dosage-level for 5-CNAC: 150 mg/kg-   Dosing volume: 5 ml/kg-   Compound concentration: 30 mg/ml    sCT/calcitonin mimetic 1:

Animals treated with oral sCT or oral calcitonin mimetic 1 receiveddoses of 1.0 mg/kg combined with 150 mg/kg 5-CNAC—all dissolved inmilliQ H20.

-   Dosage-level for sCT/calcitonin mimetic 1: 1.0 mg/kg-   Dosing volume: 5 ml/kg-   Compound concentration: 0.2 mg/ml

Drug administration by per oral (p.o.) gavage b.i.d. (7-8 am and 3-4 pm)during the study period and as single dose in the morning prior to startof OGTT.

Oral gavage of glucose during OGTT was prepared by the followingcalculation:

D-Glucose:

Animals were given 2 g/kg single dose dissolved in milliQ H20.

-   Dosage-level for D-Glucose: 2 g/kg-   Dosing volume: 4 ml/kg-   Compound concentration: 500 mg/ml

Experimental Setup:

Baseline Week 1 Week 2 Week 3 Week 4 FPG BW BW BW BW BW Food Food FoodFood B FPG FPG B B OGTT FPG = Fasting Plasma Glucose; BW = Body Weight;B = Blood; OGTT = Oral Glucose Tolerance Test

OGTT Following Overnight Fasting (16 h):

−30 0 15 30 60 120 240 min D G B B B B B B B BG BG BG BG BG BG BG D =Drug; BG = Blood glucose; B = Blood; G = Glucose

Blood sampling and glycemia were measured by heated tail venouspuncture.

Whole blood glucose levels were determined with an ACCU-CHEK® Avia bloodglucose meter (Roche Diagnostics, Rotkreuz, Switzerland). Blood (approx300 ul) is collected in 1 ml MiniCollect K3EDTA plasma-tube(Greiner-Bio-One GmbH, Frickenhausen, Germany), inverted, and stored onice. Tubes are centrifuged 3000×g (5000 rpm in table centrifuge) for 10min at 4° C. and plasma obtained. Plasma samples are stored at −20° C.until analysis. A total of ˜2.5 ml blood is obtained during OGTT (˜0.3%of body weight).

Experimental Groups

Intervention Compound Conc. Number Oral vehicle 5-CNAC 150 mg/kg n = 10Oral sCT 5-CNAC + sCT 150 mg/kg + 1 mg/kg n = 10 Oral calcitonin5-CNAC + calcitonin 150 mg/kg + 1 mg/kg n = 10 mimetic 1 mimetic 1

Statistical analysis was performed by one-way ANOVA followed by theDunnett's post hoc test for multiple comparison. Student's t-test wasperformed to compare two paired group. All analysis was performed usingGRAPHPAD PRISM software (GraphPad Prism, San Diego, Calif. U.S.A).Incremental area under curve (iAUC) during OGTT was calculated by thetrapezoidal method. A value of P<0.05 was considered to be significant.All data are presented as mean±standard error of the mean (SEM).

Results Baseline Characteristics:

Results are summarized in FIGS. 1A-1D (Food intake and body weight),FIGS. 2A-2B

(OGTT) and FIG. 3 (FPG). FIG. 1A, FIG. 1B, FIG. 1C, and FIG. 1D show theeffect of chronic oral salmon calcitonin (“sCT”) versus oral UGP 302administration on body weight and food intake in DIO rats as measured inExample 1. FIG. 2A and FIG. 2B show the effect of oral sCT versus oralUGP 302 on glucose tolerance during OGTT in DIO rats as measured inExample 1. FIG. 3 shows the effect of oral sCT versus oral UGP 302 onfasting glycemia in DIO rats as measured in Example 1;

One dose of oral sCT/calcitonin mimetic 1 containing 1 mg/kg compoundwas applied by gavage twice daily to four groups of rats for 4 weeks. Anoral vehicle group served as dosing regimen control, respectively. *P<0.05, ** P<0.01, *** P<0.001 vs Vehicle. Results are presented asmeans±SEM.

The 16-weeks ad libitum high-fat diet induced a pronounced obesephenotype in the diet-sensitive (DIO) rats when comparing body weight totheir diet-resistant (DR) littermates (P<0.001) (Table 3). 6-hrs Fastingglycemia was not different between DIO and DR. In contrast, area undercurve (AUC) calculations during OGTT was significantly higher in DIOrats compared to DR rats, demonstrating the high-fat diet-inducedglucose intolerance (Table 3).

TABLE 3 Metabolic parameters in DIO and DR rats Diet-resistant (DR)Diet-sensitive (DIO) Body Weight (g) 609.5 ± 24.5 841.8 ± 22.9***Fasting plasma glucose (mM)  6.5 ± 0.1 6.8 ± 0.2  AUC in OGTT 625.1 ±20.5 914.3 ± 44.6*** Blood glucose (mM * min) AUC, area under curve;OGTT, oral glucose tolerance test. Data are means ± SEM (n = 12/DR, n =24/DIO).

Body Weight and Food Intake

During the first week of treatment administration of oral sCTsignificantly reduced food intake compared to oral vehicle treated rats.Furthermore, oral sCT protected against further gain in body weight asobserved for oral vehicle group (FIGS. 1A-1 D). Thus, these observationsconfirm the acute strong anorectic action induced by application of oralsCT in DIO rats.

Interestingly, from week 2 of treatment and throughout the study period,food intake normalized in oral sCT treated rats and resembled ingestionby oral vehicle resulting in a lack of difference in regards tocumulative food intake at study end. This confirms previously reportssuggesting a transient effect of oral sCT upon energy intake. However,throughout the study period, oral sCT sustained the protecting effect onbody weight gain and significantly reduced body weight from baselinewhen compared to oral vehicle at study end (FIGS. 1A-1B). This is inline with a possibly endogenous effect of oral sCT upon energyexpenditure to chronically regulate energy balance.

Generally, oral application of calcitonin mimetic 1 resembled the stronganorectic action of oral sCT during the initial week of treatment andsignificantly reduced food intake and protected against gain in bodyweight compared to oral vehicle group (FIG. 1). As observed for oralsCT, calcitonin mimetic 1 exerted a transient effect on food intake,although food intake trended reduced when compared to oral sCT duringthe study period. Thus, following four weeks of treatment cumulativefood intake was significantly reduced in calcitonin mimetic 1 whencompared to oral vehicle. Furthermore, when compared to oral sCT, a morepronounced significant reduction in body weight was observed suggestingsuperiority in regards to effect on energy balance.

Glucose Tolerance:

Results are shown in FIGS. 2A-2B. One dose of oral sCT/calcitoninmimetic 1 containing 1 mg/kg compound were applied by gavage twice dailyto four groups of rats for 4 weeks. An oral vehicle group served asdosing regimen control. OGTT performed following 2 weeks of treatmentafter overnight-fasting.*** P<0.001 vs Vehicle. Results are presented asmeans±SEM.

Oral sCT significantly reduced glucose iAUC during OGTT after 2 weeks oftreatment compared to oral vehicle, thus confirming the postprandialglycemic control exerted by oral application of sCT as previouslydemonstrated. In general, calcitonin mimetic 1 demonstrated a similarsignificant reduction in iAUC as observed for oral sCT, although with noclear superiority to oral sCT in this respect.

Fasting glycaemia:

Following 2 and 4 weeks of treatment, oral sCT application was notsignificantly different from oral vehicle treated rats, which is incontrast with previously observations in male DIO rats, in where a 1-1.5mM reduction in fasting blood glucose typically is observed followingchronic treatment. For calcitonin mimetic 1, a trend towards superiorityin fasting glycaemia was observed throughout the study period whencompared to oral vehicle or oral sCT.

EXAMPLE 2 Acute and Short Term Effects of Oral sCT Versus OralCalcitonin Mimetic 1 Animals

The study was performed in male Levin-DIO rats (diet-sensitive) andLevin-DR (diet-resistant) (TacLevin: CD (SD) DIO) (Taconic, Hudson,N.Y., U.S.A.) obtained at age 6-7 weeks. On arrival, DIO rats were givenhigh fat diet (60 kcal %) (#D12495, Research Diets Inc., New Brunswick,N.J., USA) and kept on the same diet for 12 weeks prior to and duringthe experiment. DR rats were given low-fat diet and served as controlgroup. Animals were pair-wise housed throughout the study. Rats werehandled and pre-dosed once daily with MilliQ H2O for 2-3 weeks prior toexperimental start to reduce stress-induced hyperglycaemia. On the dayprior to study start animals were given a single dose of vehicle.Baseline parameters were recorded in an overnight fasting (16 h)condition. Rats were randomized into treatment groups based on fastingbody weight (BW) and fasting plasma glucose (FPG). Body weight, food andwater intake were recorded prior to and at study end.

Compounds

Oral sCT/calcitonin mimetic 1 solution was prepared on the day of dosingby mixing the carrier with the given compound based on the followingcalculations:

5-CNAC (vehicle):

Animals treated with oral 5-CNAC received a dose of 150 mg/kg dissolvedin milliQ H20.

-   Dosage-level for 5-CNAC: 150 mg/kg-   Dosing volume: 5 ml/kg-   Compound concentration: 30 mg/ml    sCT/calcitonin mimetic 1:

Animals treated with oral sCT or oral calcitonin mimetic 1 receiveddoses of 0.5 mg/kg, 1.0 mg/kg or 2.0 mg/kg combined with 150 mg/kg5-CNAC—all dissolved in milliQ H20.

-   Dosage-level for sCT/calcitonin mimetic 1:0.5 mg/kg-   Dosing volume: 5 ml/kg-   Compound concentration: 0.1 mg/ml-   Dosage-level for sCT/calcitonin mimetic 1:1.0 mg/kg-   Dosing volume: 5 ml/kg-   Compound concentration: 0.2 mg/ml-   Dosage-level for sCT/calcitonin mimetic 1:2.0 mg/kg-   Dosing volume: 5 ml/kg-   Compound concentration: 0.4 mg/ml

Drug administration were given by per oral (p.o.) gavage b.i.d. duringthe study period and as single dose in the morning prior to start ofOGTT.

Oral gavage of glucose during OGTT was prepared by the followingcalculation:

D-Glucose:

Animals were given 2 g/kg single dose dissolved in milliQ H20.

-   Dosage-level for D-Glucose: 2 g/kg

Dosing volume: 4 ml/kg

-   Compound concentration: 500 mg/ml

Experimental Setup

Acute Testing—Treatment Period for 0.5 mg/kg, 1 mg/kg and 2 mg/kg:

Day 0 Day 1-2 Day 3 Day 4 Day 5 Day 6 Day 7 1^(st) OGTT Rest Pre-doseTreatment Treatment Treatment 2^(nd) OGTT All vehicle No All vehicle(b.i.d) (b.i.d) (b.i.d) Single handling dose

Following the initial (1^(st)) OGTT, animals are randomized intotreatment groups based on FBG and BW. Animals will be pre-treated 3 days(b.i.d.) prior to 2^(nd) OGTT. Dosing will be performed in the morning(7-8 am) and afternoon (3-4 pm).

The study was performed in an x-over design with each animal being itsown control.

OGTT Following Overnight Fasting (16 h):

−30 0 15 30 60 120 240 min D G B B B B B B B BG BG BG BG BG BG BG D =Drug; BG = Blood glucose; B = Blood; G = Glucose

Blood sampling and glycemia were measured by heated tail venouspuncture.

Whole blood glucose levels were determined with an ACCU-CHEK® Avia bloodglucose meter (Roche Diagnostics, Rotkreuz, Switzerland). Blood (approx300 ul) is collected in 1 ml MiniCollect K3EDTA plasma-tube(Greiner-Bio-One GmbH, Frickenhausen, Germany), inverted, and stored onice. Tubes are centrifuged 3000×g (5000 rpm in table centrifuge) for 10min at 4° C. and plasma obtained. Plasma samples are stored at −20° C.until analysis. A total of ˜2.5 ml blood is obtained during OGTT (˜0.3%of body weight).

Experimental Groups

Intervention Compound Conc. Number Oral vehicle 5-CNAC 150 mg/kg (4groups of n = 8) X-over design to 0.5 mg/kg Oral sCT 5-CNAC + sCT 150mg/kg + n = 8 0.5 mg/kg Oral calcitonin 5-CNAC + calcitonin 150 mg/kg n= 8 mimetic 1 mimetic 1 0.5 mg/kg Oral vehicle 5-CNAC 150 mg/kg (4groups of n = 8) X-over design to 1 mg/kg Oral sCT 5-CNAC + sCT 150mg/kg + n = 8 1 mg/kg Oral calcitonin 5-CNAC + calcitonin 150 mg/kg n =8 mimetic 1 mimetic 1 1 mg/kg Oral vehicle 5-CNAC 150 mg/kg (4 groups ofn = 8) X-over design to 2 mg/kg Oral sCT 5-CNAC + sCT 150 mg/kg + n = 82 mg/kg Oral calcitonin 5-CNAC + calcitonin 150 mg/kg n = 8 mimetic 1mimetic 1 2 mg/kg

Statistics

Statistical analysis was performed by one-way ANOVA followed by theDunnett's post hoc test for multiple comparison. Student's t-test wasperformed to compare two paired group. All analysis was performed usingGRAPHPAD PRISM software (GraphPad Prism, San Diego, Calif. U.S.A).Incremental area under curve (iAUC) during OGTT was calculated by thetrapezoidal method. A value of P<0.05 was considered to be significant.All data are presented as mean±standard error of the mean (SEM).

Results Baseline Characteristics

The 12-weeks ad libitum high-fat diet induced a pronounced obesephenotype in the diet-sensitive (DIO) rats when comparing body weight totheir diet-resistant (DR) littermates (P<0.001) (Table 1). Fastingglycemia was not different between DIO and DR. In contrast, area undercurve (AUC) calculations during OGTT was significantly higher in DIOrats compared to DR rats, demonstrating the high-fat diet-inducedglucose intolerance (Table 4).

TABLE 4 Metabolic parameters in DIO and DR rats Diet-resistant (DR)Diet-sensitive (DIO) Body Weight (g) 609.5 ± 24.5 813.6 ± 9.8*** Fasting plasma glucose (mM)  5.8 ± 0.1 5.8 ± 0.2  AUC in OGTT 648.8 ±27.3 888.4 ± 64.3*** Blood glucose (mM * min) AUC, area under curve;OGTT, oral glucose tolerance test. Data are means ± SEM (n = 12/DR, n =24/DIO).

Body Weight and Food Intake

Three different doses of oral sCT/calcitonin mimetic 1 containing 0.5, 1and 2 mg/kg compound were applied by gavage twice daily to 4 groups ofrats for 3 days. * P<0.05, ** P<0.01 vs oral sCT.

Results are presented in FIGS. 4A-4B, FIGS. 5A-5B, and FIGS. 6A-6B asmeans±SEM. FIG. 4A and FIG. 4B show the effect of oral sCT versus oralUGP 302 on body weight and food intake in DIO rats observed in Example 2at a first dosage. FIG. 5A and FIG. 5B show the effect of oral sCTversus oral UGP 302 on body weight and food intake in DIO rats observedin Example 2 at a second dosage. FIG. 6A and FIG. 6B show the effect oforal sCT versus oral UGP 302 on body weight and food intake in DIO ratsobserved in Example 2 at a third dosage;

Oral sCT dose-dependently decreased body weight and food intakefollowing the short-term treatment period and thus confirmed theanorectic action induced by targeting the amylin receptor as previouslyobserved. In general, the mimetic demonstrated dose-dependentsuperiority to oral sCT in regards to reduction in body weight asillustrated in FIG. 4A-4B, FIG. 5A-5B and FIG. 6A-6B. Application ofcalcitonin mimetic 1 at 0.5 mg/kg demonstrated significantly differenceto oral sCT 0.5 mg/kg. The food intake for the mimetic trendeddose-dependently reduced compared to oral sCT.

Glucose Tolerance

Three different doses of oral sCT/calcitonin mimetic 1 containing 0.5, 1and 2 mg/kg compound were applied by gavage twice daily to 4 groups ofrats for 3 days prior to OGTT. The experimental set-up was a cross-overdesign. *P<0.05, ** P<0.01, *** P<0.001 vs oral vehicle. Results arepresented in FIG. 7A-7B and FIG. 8A-8B as means±SEM. FIG. 7A and FIG. 7Bshow the effect of oral sCT versus oral UGP 302 at a first dosage onglucose tolerance during OGTT in DIO rats as measured in Example 2. FIG.8A and FIG. 8B show the effect of oral sCT versus oral UGP 302 at asecond dosage on glucose tolerance during OGTT in DIO rats as measuredin Example 2.

Oral sCT significantly reduced glucose iAUC during OGTT for 0.5, 1 and 2mg/kg doses compared to oral vehicle, thus confirming the postprandialglycemic control exerted by oral application of sCT as previouslydemonstrated. Calcitonin mimetic 1 demonstrated a similar significantlyreduction in iAUC as observed for oral sCT, although with no clearsuperiority to oral sCT within the various UGPs.

EXAMPLE 3 Acute and Short Term Effects of Oral sCT Versus UGP284, UGP298and UGP302 Animals

The study was performed in male Levin-DIO rats (diet-sensitive) andLevin-DR (diet-resistant) (TacLevin: CD (SD) DIO) (Taconic, Hudson,N.Y., U.S.A.) obtained at age 6-7 weeks. On arrival, DIO rats were givenhigh fat diet (60 kcal %) (#D12495, Research Diets Inc., New Brunswick,N.J., USA) and kept on the same diet for 12 weeks prior to and duringthe experiment. DR rats were given low-fat diet and served as controlgroup. Animals were pair-wise housed throughout the study. Rats werehandled and pre-dosed once daily with MilliQ H2O for 2-3 weeks prior toexperimental start to reduce stress-induced hyperglycemia. On the dayprior to study start animals were given a single dose of vehicle.Baseline parameters were recorded in an overnight fasting (16 h)condition. Rats were randomized into treatment groups based on fastingbody weight (BW) and fasting plasma glucose (FPG). Body weight, food andwater intake were recorded prior to and at study end.

Compound

Oral sCT/UGP solution was prepared on the day of dosing by mixing thecarrier with the given compound based on the following calculations:

5-CNAC (Vehicle):

Animals treated with oral 5-CNAC received a dose of 150 mg/kg dissolvedin milliQ H20.

-   Dosage-level for 5-CNAC: 150 mg/kg-   Dosing volume: 5 ml/kg-   Compound concentration: 30 mg/ml    (sCT/UGP284/UGP298/UGP302)

Animals treated with oral sCT or oral UGP284/UGP298/UGP302 receiveddoses of 0.5 mg/kg, 1.0 mg/kg or 2.0 mg/kg combined with 150 mg/kg5-CNAC—all dissolved in milliQ H₂O.

-   Dosage-level for sCT/UGP284/UGP298/UGP302: 0.5 mg/kg-   Dosing volume: 5 ml/kg-   Compound concentration: 0.1 mg/ml-   Dosage-level for sCT/UGP284/UGP298/UGP302: 1.0 mg/kg-   Dosing volume: 5 ml/kg-   Compound concentration: 0.2 mg/ml

Dosage-level for sCT/UGP284/UGP298/UGP302: 2.0 mg/kg

-   Dosing volume: 5 ml/kg-   Compound concentration: 0.4 mg/ml

Drug administration were given by per oral (p.o.) gavage b.i.d. duringthe study period and as single dose in the morning prior to start ofOGTT.

Oral gavage of glucose during OGTT was prepared by the followingcalculation: D-Glucose:

-   Animals were given 2 g/kg single dose dissolved in milliQ H20.-   Dosage-level for D-Glucose: 2 g/kg-   Dosing volume: 4 ml/kg-   Compound concentration: 500 mg/ml

Experimental Setup

Acute Testing—Treatment Period for 0.5 mg/kg, 1 mg/kg and 2 mg/kg:

Day 0 Day 1-2 Day 3 Day 4 Day 5 Day 6 Day 7 1^(st) OGTT Rest Pre-doseTreatment Treatment Treatment 2^(nd) OGTT All vehicle No handling Allvehicle (b.i.d) (b.i.d) (b.i.d) Single dose

Following the initial (1^(st)) OGTT, animals were randomized intotreatment groups based on FBG and BW. Animals were pre-treated 3 days(b.i.d.) prior to 2^(nd) OGTT. The study was performed in an x-overdesign with each animal being its own control.

OGTT Following Overnight Fasting (16 h):

−30 0 15 30 60 120 240 min D G B B B B B B B BG BG BG BG BG BG BG D =Drug; BG = Blood glucose; B = Blood; G = Glucose

Blood sampling and glycemia were measured by heated tail venouspuncture. Whole blood glucose levels were determined with an ACCU-CHEK®Avia blood glucose meter (Roche Diagnostics, Rotkreuz, Switzerland).Blood (approx 300 μl) is collected in 1 ml MiniCollect K3EDTAplasma-tube (Greiner-Bio-One GmbH, Frickenhausen, Germany), inverted,and stored on ice. Tubes are centrifuged 3000×g (5000 rpm in tablecentrifuge) for 10 min at 4° C. and plasma obtained. Plasma samples arestored at −20° C. until analysis. A total of ˜2.5 ml blood is obtainedduring OGTT (˜0.3% of body weight).

Experimental Groups:

Intervention Compound Conc. Number Oral vehicle 5-CNAC 150 mg/kg (4groups of n = 8) X-over design to 0.5 mg/kg Oral sCT 5-CNAC + 150mg/kg + n = 8 sCT 0.5 mg/kg Oral UGP284 5-CNAC + 150 mg/kg n = 8 UGP2840.5 mg/kg Oral UGP298 5-CNAC + 150 mg/kg n = 8 UGP298 0.5 mg/kg OralUGP302 5-CNAC + 150 mg/kg n = 8 UGP302 0.5 mg/kg Oral vehicle 5-CNAC 150mg/kg (4 groups of n = 8) X-over design to 1 mg/kg Oral sCT 5-CNAC + 150mg/kg + n = 8 sCT 1 mg/kg Oral UGP284 5-CNAC + 150 mg/kg n = 8 UGP284 1mg/kg Oral UGP298 5-CNAC + 150 mg/kg n = 8 UGP298 1 mg/kg Oral UGP3025-CNAC + 150 mg/kg n = 8 UGP302 1 mg/kg Oral vehicle 5-CNAC 150 mg/kg (4groups of n = 8) X-over design to 2 mg/kg Oral sCT 5-CNAC + 150 mg/kg +n = 8 sCT 2 mg/kg Oral UGP284 5-CNAC + 150 mg/kg n = 8 UGP284 2 mg/kgOral UGP298 5-CNAC + 150 mg/kg n = 8 UGP298 2 mg/kg Oral UGP302 5-CNAC +150 mg/kg n = 8 UGP302 2 mg/kg

Statistics

Statistical analysis was performed by one-way ANOVA followed by theDunnett's post hoc test for multiple comparison. Student's t-test wasperformed to compare two paired group. All analysis was performed usingGRAPHPAD PRISM software (GraphPad Prism, San Diego, Calif. U.S.A).Incremental area under curve (iAUC) during OGTT was calculated by thetrapezoidal method. A value of P<0.05 was considered to be significant.All data are presented as mean±standard error of the mean (SEM).

Results Baseline Characteristics

The 12-weeks ad libitum high-fat diet induced a pronounced obesephenotype in the diet-sensitive (DIO) rats when comparing body weight totheir diet-resistant (DR) littermates (P<0.001) (Table 5). Fastingglycemia was not different between DIO and DR. In contrast, area undercurve (AUC) calculations during OGTT were significantly higher in DIOrats compared to DR rats, demonstrating the high-fat diet-inducedglucose intolerance (Table 5).

TABLE 5 Metabolic parameters in DIO and DR rats Diet-resistant (DR)Diet-sensitive (DIO) Body Weight (g) 609.5 ± 24.5 813.6 ± 9.8*** Fasting plasma glucose (mM)  5.8 ± 0.1 5.8 ± 0.2  AUC in OGTT 648.8 ±27.3 888.4 ± 64.3*** Blood glucose (mM * min) AUC, area under curve;OGTT, oral glucose tolerance test. Data are means ± SEM (n = 12/DR, n =24/DIO).

Body Weight and Food Intake

Oral sCT dose-dependently decreased body weight and food intakefollowing the short-term treatment period and thus confirmed theanorectic action induced by targeting the amylin receptor as previouslyobserved. In general, all UGP mimetics demonstrated dose-dependentlysuperiority to oral sCT in regards to reduction in body weight asillustrated in FIGS. 9A-9F. Application of UGP302 at 0.5 mg/kgdemonstrated significantly difference to oral sCT 0.5 mg/kg. For UGP284,significantly difference at 2 mg/kg dose was observed when compared tooral sCT 2 mg/kg. Finally, UGP298 at both 1 mg/kg and 2 mg/kg doses weresignificantly different when compared with oral sCT at similar doses.FIG. 9A, FIG. 9B, FIG. 9C, FIG. 9D, FIG. 9E, and FIG. 9F show the effectof three different doses of oral sCT/UGP284/UGP298/UGP302 containing0.5, 1 and 2 mg/kg compound were applied by gavage twice daily to 4groups of rats for 3 days. * P<0.05, ** P<0.01 vs oral sCT. Results arepresented as means±SEM.

Glucose Tolerance

FIG. 10A, FIG. 10B, FIG. 10C, FIG. 10D, FIG. 10E, and FIG. 10F show theeffect of oral sCT versus oral UGPs on glucose tolerance during OGTT inDIO rats. Three different doses of oral sCT/UGP284/UGP298/UGP302containing 0.5, 1 and 2 mg/kg compound were applied by gavage twicedaily to 4 groups of rats for 3 days prior to OGTT. The experimentalset-up was a cross-over design. * P<0.05, ** P<0.01, *** P<0.001 vs oralvehicle. Results are presented as means±SEM. All UGPs demonstrated asimilar significant reduction in iAUC as observed for oral sCT.

In conclusion, application of UGP284, UGP298 and UGP302 at 0.5, 1 and 2mg/kg doses demonstrated superiority to equivalent doses of oral sCT inregards to energy balance in male DIO rats. Furthermore, UGP284, UGP298and UGP302 at doses of 0.5, 1 and 2 mg/kg produced an improvement inglucose tolerance during OGTT.

EXAMPLE 4 Binding of sCT Analogs to T47D Cell Calcitonin Receptors

sCT analogs at various concentrations were tested in a T47D (humanbreast epithelial cell line) bioassay. This cell line is known to havethe following receptors: calcitonin, androgen, progesterone,glucocrticoid, prolactin and estrogen. The results are presented in FIG.11 as % cAMP binding relative to sCT which was set at 100% cAMP bindingat a concentration of 1000 pg/mL. It can be seen that UGP302 providesthe highest level of binding of all the tested compounds and that itprovides a higher level of binding than sCT.

EXAMPLE 5 Food Consumption and Weight Change in Rats Fed sCT Analogs

Male Sprague-Dawley rats were housed individually in cages in which thelight/dark cycle was reversed. Rats were allowed to eat ad libitum. Foodconsumption and rat weights were monitored daily during each study. Ratswere injected intramuscularly with a saline placebo or the indicatedpeptide at the specified dose in saline. The data in the followingtables is summarized as the mean change in food consumption relative tothe day before treatment began (day -1) and as the mean change in weightrelative to the day before treatment began.

The results are shown in FIGS. 12A-12B, 13A-13B, 14A-14B, 15A-15B,16A-16B, and 17A-17B. FIG. 12A and FIG. 12B show food consumption andweight change measurements, respectively, for UGP 282 as measured inExample 5. FIG. 13A and FIG. 13B show food consumption and weight changemeasurements, respectively, for UGP 283 as measured in Example 5. FIG.14A and FIG. 14B show food consumption and weight change measurements,respectively, for UGP 284 as measured in Example 5. FIG. 15A and FIG.15B show food consumption and weight change measurements, respectively,for UGP 298 as measured in Example 5. FIG. 16A and FIG. 16B show foodconsumption and weight change measurements, respectively, for UGP 302 asmeasured in Example 5. FIG. 17A and FIG. 17B show food consumption andweight change measurements, respectively, for UGP 303 as measured inExample 5.

It can be seen that all of the tested compounds induce weight loss andreduce feed intake.

EXAMPLE 6 Markers of Osteoporosis and Osteoarthritis

The effect of sCT/calcitonin mimetic on bone and cartilage loss wasstudied in DIO rats. The animals were dosed as described in Table 6below, and 2 hours after treatment blood sampling was done by heatedtail venous puncture.

Serum CTX-I levels, as an indication of bone resorption, were measuredusing the RatLaps™ ELISA, and serum CTX-II levels, as an indication ofcartilage degradation, were measured using the Serum PC Cartilaps™ELISA.

TABLE 6 Experimental groups Intervention Compound Conc. Number Oralvehicle 5-CNAC 150 mg/kg n = 8 Oral sCT 5-CNAC + sCT 150 mg/kg + 1 mg/kgn = 8 Oral calcitonin 5-CNAC + 150 mg/kg + 1 mg/kg n = 8 mimetic of SEQID NO: 15 SEQ ID NO: 18

The results are seen in FIG. 18 and FIG. 19, where a calcitonin mimeticof SEQ ID NO: 18 shows a stronger effect in reduction of both boneresorption and cartilage degradation than does sCT.

In some embodiments, a peptide of the present disclosure has a sequenceselected from SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO:14, SEQ ID NO: 15, SEQ ID NO: 17 and SEQ ID NO: 18.

In some embodiments, a method includes administering to a patient aneffective amount of a peptide selected from the group consisting of: SEQID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15,and SEQ ID NO: 17 to affect a weight reduction in the patient.

In some embodiments, a method includes administering to a patient aneffective amount of a peptide selected from the group consisting of: SEQID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15,and SEQ ID NO: 17 to affect postprandial glycemic control in thepatient.

In some embodiments, a method includes administering to the patient aneffective amount of a peptide selected from the group consisting of SEQID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15,and SEQ ID NO: 17 to affect an improvement in glycemic control in thepatient.

In some embodiments, a method includes administering to a patient aneffective amount of a peptide of SEQ ID NO: 18 having the sequenceC_(m)SNLSTCVLGKLSQELH KLQTYPRTDVGANXaaXaa_(a) so as to reduce at leastone of bone resorption and cartilage degradation in the patient.

EXAMPLE 7 Combination Treatment with Metformin

To assess the potential of UGP302 (SEQ ID NO: 15) in combination withmetformin,

Zucker Diabetic Fatty rats were used. At the age of 6 weeks 40 rats wererandomized into four groups according to HbA1c, FPG and BW. The groupswere: Control (Vehicle), Metformin 400 mg/kg/day, UGP302 5 μg/kg/day andthe combination of the two molecules in the stated respective amounts.The treatments were given once daily, and metformin was dosed orally,while UGP302 was given subcutaneously, and the animals were treated for8 weeks. Fasting blood glucose (FPG) and non-fasted blood glucose (PPG)were monitored biweekly, while HbA1c was measured at termination.

As can be seen in FIGS. 20, 21 and 22, both metformin and UGP302 alonereduced both FPG and PPG, with UGP302 providing a pronounced reductionthroughout the study, while metformin lost the ability to control bloodglucose with time. In the combination group, a markedly superiorresponse was observed in both FPG and PPG, clearly showing that thesemolecules work as adjunct therapies. This synergistic effect is alsoseen in the HbA1c levels, where the combination leads to a superiorimprovement when compared to both vehicle and either molecule alone.Particularly notably, the superior effect of the combination as comparedto UGP302 alone persisted throughout the test, even though theindividual effect of metformin did not.

To further assess the effect of both single doses of each molecule andthe combination of these at those same doses, the rats underwent an oralglucose tolerance test (OGTT, FIGS. 23A-23B) and an insulin tolerancetest (ITT, FIGS. 24A-24B). In these tests the activity of both moleculesis apparent, as they individually improve both glucose and insulintolerance.

However, the combination of UGP302 and metformin was superior to eithermolecule alone, both in terms of glucose tolerance and insulin tolerance(FIGS. 23A-23B), clearly indicating the benefit of the combination.

It will be appreciated that several of the above-disclosed and otherfeatures and functions, or alternatives thereof, may be desirablycombined into many other different systems or applications. Variouspresently unforeseen or unanticipated alternatives, modifications,variations, or improvements therein may be subsequently made by thoseskilled in the art which are also intended to be encompassed by thefollowing claims.

What is claimed is:
 1. A method for treating type I diabetes, type IIdiabetes or a metabolic syndrome, obesity, or for appetite suppression,or for mitigating insulin resistance, or for reducing an undesirablyhigh fasting serum glucose level, or for reducing an undesirably highpeak serum glucose level, or for reducing an undesirably high peak seruminsulin level, or for reducing an undesirably large response to aglucose tolerance test, or for treating osteoporosis, or for treatingosteoarthritis, comprising administration to the patient an effectiveamount of a peptide selected from the group consisting of:AcCSNLSTCVLGRLSQELHRLQTFPRTDVGA SEQ ID NO: 12 NTAcYAcCSNLSTCVLGKLSQELHKLQTYPRTDVGA SEQ ID NO: 15 NAP-NH₂SuccCSNLSTCVLGKLSQELHKLQTYPRTDV SEQ ID NO: 17 GANAY-NH₂


2. The method of claim 1, wherein method is for treating type Idiabetes, type II diabetes, or a metabolic syndrome.
 3. The method ofclaim 1, wherein the method is for treating obesity
 4. The method ofclaim 1, wherein the method is for appetite suppression.
 5. The methodof claim 1, wherein the method is for mitigating insulin resistance. 6.The method of claim 1, wherein the method is for reducing an undesirablyhigh fasting serum glucose level, or for reducing an undesirably highpeak serum glucose level, or for reducing an undesirably high peak seruminsulin level, or for reducing an undesirably large response to aglucose tolerance test.
 7. The method of claim 1, wherein the method isfor treating osteoporosis or osteoarthritis.
 8. The method of claim 1,wherein the peptide is: AcCSNLSTCVLGKLSQELHKLQTYPRTDVGA SEQ ID NO: 15NAP-NH₂


9. The method of claim 1, wherein the peptide is formulated for enteraladministration.
 10. The method of claim 1, wherein the peptide isformulated for parenteral administration.
 11. The method of claim 1,wherein the peptide is formulated with a carrier for oraladministration.
 12. The method of claim 11, wherein the carriercomprises N-(5-chlorosalicyloyl)-8-aminocaprylic acid (5-CNAC), sodiumsalt of 10-(2-Hydroxybenzamido)decanoic acid (SNAD), or sodium salt ofN-(8-[2-hydroxybenzoyl]amino)caprylic acid (SNAC).
 13. The method ofclaim 1, wherein the peptide is formulated in a pharmaceuticalcomposition for oral administration comprising coated citric acidparticles, and wherein the coated citric acid particles increases theoral bioavailability of the peptide.